[Objective] The research aimed to reveal the functions of NAC and UBA domains in Peatl＇s thermal stability. [Method] Fusion expression vectors of Pearl protein and the 3 deletion mutants were constructed. The recombi...[Objective] The research aimed to reveal the functions of NAC and UBA domains in Peatl＇s thermal stability. [Method] Fusion expression vectors of Pearl protein and the 3 deletion mutants were constructed. The recombinant plasmids were induced by IPTG and the target proteins （Peatl, Peatl-△CD99,Peatl-△ND49 and Pearl-△ND108 ）were expressed obtained by AKTA and its thermal stability was analyzed. [Result] The research found that 3 deletion mutants have good thermal stability like Pearl. [Conclusion] The research demonstrated that the coexistence of NAC or UBA domains is not necessary to thermal stability of Pearl protein , and they may give the protein particular stability structure seperately.展开更多
The peaT1 gene fragment was amplified from pGEM-6p-l-peaT1 by PCR, and recovered target gene was cloned into pLexA vector. After digestion and sequencing, the bait vector pLexA-peaT1 was transformed into yeast strain ...The peaT1 gene fragment was amplified from pGEM-6p-l-peaT1 by PCR, and recovered target gene was cloned into pLexA vector. After digestion and sequencing, the bait vector pLexA-peaT1 was transformed into yeast strain EGY48 [p8op-lacZ] by PEG/LiAC, and the transcriptional activity of bait vector was detected. The results showed that recombinant bait plasmid pLexA-PEMG1 was constructed, for the two bands of recombinant bait plasmid in agarose gel eleetrophoresis were expected after digesting by restriction endonuclease EcoR I and Xho I. Therefore, the recombinant bait plasmid could be used in yeast two-hybrid system to screen a cDNA library.展开更多
基金Supported by the“973”Program(2003CB114204)the Science and Technology Plan(D0706005040431)~~
文摘[Objective] The research aimed to reveal the functions of NAC and UBA domains in Peatl＇s thermal stability. [Method] Fusion expression vectors of Pearl protein and the 3 deletion mutants were constructed. The recombinant plasmids were induced by IPTG and the target proteins （Peatl, Peatl-△CD99,Peatl-△ND49 and Pearl-△ND108 ）were expressed obtained by AKTA and its thermal stability was analyzed. [Result] The research found that 3 deletion mutants have good thermal stability like Pearl. [Conclusion] The research demonstrated that the coexistence of NAC or UBA domains is not necessary to thermal stability of Pearl protein , and they may give the protein particular stability structure seperately.
基金Supported by the National“863”Program(2006AA10A210)~~
文摘The peaT1 gene fragment was amplified from pGEM-6p-l-peaT1 by PCR, and recovered target gene was cloned into pLexA vector. After digestion and sequencing, the bait vector pLexA-peaT1 was transformed into yeast strain EGY48 [p8op-lacZ] by PEG/LiAC, and the transcriptional activity of bait vector was detected. The results showed that recombinant bait plasmid pLexA-PEMG1 was constructed, for the two bands of recombinant bait plasmid in agarose gel eleetrophoresis were expected after digesting by restriction endonuclease EcoR I and Xho I. Therefore, the recombinant bait plasmid could be used in yeast two-hybrid system to screen a cDNA library.
